Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Expression of the renal Na+/dicarboxylate cotransporter, NaDC-1, in COS-7 cells

  • 50 Accesses

  • 11 Citations

Abstract

The cloned rabbit renal Na+/dicarboxylate cotransporter, NaDC-1, was transiently expressed in the mammalian monkey kidney cell line, COS-7. Cells transfected with the plasmid pSV-201, containing the cDNA for the Na+/dicarboxylate cotransporter, expressed sodium-dependent succinate and citrate transport after 48 h. Cells transfected with control plasmid, pSV-SPORT, did not express sodium-dependent transport of succinate or citrate. The transport of succinate in cells expressing NaDC-1 was inhibited by di-and tricarboxylic acids, but not by monocarboxylic acids. Sodium-dependent transport of succinate was insensitive to changes in pH, while sodium-dependent citrate transport was stimulated by acidic pH. Succinate transport by NaDC-1 was saturable with an apparent Michaelis constant,K m, around 0.5 mM. The kinetics of sodium activation of succinate transport by NaDC-1 were sigmoidal, with an apparent Hill coefficient of 2.9, indicating that three sodium ions are involved in the transport of each succinate. Succinate transport by NaDC-1 was inhibited by lithium. The functional characteristics of NaDC-1 expressed in COS-7 cells correspond to those of the Na+/dicarboxylate cotransporter of the apical membrane of the renal proximal tubule.

This is a preview of subscription content, log in to check access.

References

  1. 1.

    Birnir B, Lee HS, Hediger MA, Wright EM (1990) Expression and characterization of the intestinal Na+/glucose cotransporter in COS-7 cells. Biochim Biophys Acta 1048:100–104

  2. 2.

    Brennan S, Hering-Smith K, Hamm LL (1988) Effect of pH on citrate reabsorption in the proximal convoluted tubule. Am J Physiol 255:F301-F306

  3. 3.

    Burckhardt G (1984) Sodium-dependent dicarboxylate transport in rat renal basolateral membrane vesicles. Pflügers Arch 401:254–261

  4. 4.

    Busch AE, Waldegger S, Herzer T, Biber J, Markovich D, Murer H, Lang F (1994) Electrogenic cotransport of Na+ and sulfate in Xenopus oocytes expressing the cloned Na+/SO4 4 2−2 -transport protein NaSi-1. J Biol Chem 269:12407–12409

  5. 5.

    Domin BA, Mahony WB, Zimmerman TP (1988) Purine nucleobase transport in human erythrocytes. J Biol Chem 263: 9276–9284

  6. 6.

    Hamm LL (1990) Renal handling of citrate. Kidney Int 38:728–735

  7. 7.

    Hirayama B, Wright EM (1986) Coupling between sodium and succinate transport across renal brush border membrane vesicles. Pflügers Arch 407 [Suppl 2]:S174-S179

  8. 8.

    Jenkins AD, Dousa TP, Smith LH (1985) Transport of citrate across renal brush border membrane: effects of dietary acid and alkali loading. Am J Physiol 249: F590-F595

  9. 9.

    Levi M, McDonald LA, Preisig PA, Alpern RJ (1991) Chronic K depletion stimulates rat renal brush-border membrane Nacitrate cotransporter. Am J Physiol 261:F767-F773

  10. 10.

    Markovich D, Forgo J, Stange G, Biber J, Murer H (1993) Expression cloning of rat renal Na+/SO 4 2− - cotransport. Proc Natl Acad Sci USA 90:8073–8077

  11. 11.

    Pajor AM (1995) Sequence and functional characterization of a renal sodium/ dicarboxylate cotransporter. J Biol Chem 270: 5779–5785

  12. 12.

    Pak CYC (1991) Etiology and treatment of urolithiasis. Am J Kidney Dis 18:624–637

  13. 13.

    Sigel E (1990) Use ofXenopus oocytes for the functional expression of plasma membrane proteins. J Membr Biol 117: 201–221

  14. 14.

    Smith CD, Hirayama BA, Wright EM (1992) Baculovirus-mediated expression of the Na+/glucose cotransporter in Sf9 cells. Biochim Biophys Acta 104:151–159

  15. 15.

    Windus DW, Cohn DE, Heifets M (1986) Effects of fasting on citrate transport by the brush-border membrane of the rat kidney. Am J Physiol 251:F678-F682

  16. 16.

    Wright EM (1985) Transport of carboxylic acids by renal membrane vesicles. Annu Rev Physiol 47:127–141

  17. 17.

    Wright SH, Wunz TM (1987) Succinate and citrate transport in renal basolateral and brush-border membranes. Am J Physiol 253:F432-F439

  18. 18.

    Wright SH, Kippen I, Wright EM (1982) Effect of pH on the transport of Krebs cycle intermediates in renal brush border membranes. Biochim Biophys Acta 684:287–290

  19. 19.

    Wright SH, Hirayama B, Kaunitz JD, Kippen I, Wright EM (1983) Kinetics of sodium succinate cotransport across renal brush-border membranes. J Biol Chem 258:5456–5462

Download references

Author information

Correspondence to Ana M. Pajor.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Pajor, A.M., Valmonte, H.G. Expression of the renal Na+/dicarboxylate cotransporter, NaDC-1, in COS-7 cells. Pflugers Arch. 431, 645–651 (1996). https://doi.org/10.1007/BF02191915

Download citation

Key words

  • Na+/succinate cotransporter
  • Expression in mammalian cell
  • Succinate transporter
  • COS-7 cells